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为观察猪流行性腹泻病毒(PDEV)的血凝特性,进而为建立PEDV HA和HI试验,评估疫苗免疫抗体水平奠定基础,将CV777、DR13、YN13和YN144共4株PEDV毒株,在37 ℃下,分别用0、5、10、50、100 μg/mL胰蛋白酶处理30 min,然后用鸡、兔和猪红细胞进行血凝(HA)试验;用已经鉴定为PEDV抗体阳性的血清和乳清分别进行血凝抑制(HI)试验。结果显示:4株PEDV毒株只与兔红细胞发生凝集反应;CV777、DR13毒株的HA现象需要用5 μg/mL的胰蛋白酶进行预处理,但是YN13和YN144毒株的HA现象不需要胰蛋白酶作用;该凝集现象能够被PEDV抗体特异性抑制。结果表明,PEDV能够凝集兔的红细胞,不能凝集鸡和猪的红细胞,因而可以用兔红细胞建立PEDV的HA和HI试验方法。
HA-HI Test for Four Strains of Porcine Epidemic Diarrhea Virus
In order to observe the hemagglutination(HA)of porcine epidemic diarrhea virus(PDEV)and to lay a foundation for development of HA-HI test for PEDV and evaluation of the antibody level,four strains of PEDV,including CV777,DR13,YN13 and YN144,were treated with trypsin of 0,5,10,50 and 100 μg/mL respectively for 30 minutes at 37 ℃,followed by HA test by use of red blood cells(RBC)of chicken,rabbit and pig respectively;then hemagglutination inhibition(HI)test was carried out with serum and whey that had been identified to be PEDV-positive. The results showed that four strains of PEDV only agglutinated with rabbit RBC. After CV777 and DR13 strain were pretreated with 5 μg/mL trypsin,the HA phenomenon occurred,while no trypsin was needed for YN13 andYN144 strain. In addition,HA could be specifically inhibited by antibody against PEDV. In conclusion,rabbit RBC rather than chicken and swine RBC could be agglutinated by PEDV,therefore,the HA-HI test for PEDV could be developed by use of rabbit RBC.
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